pGLO Bacterial Transformation Lab

[@ginareyes6762]

This is the best explanation for this lab! Makes so much sense and helps visually! Thank you so much Doc Ron Bio.

[@vspianist]

I have a strong feeling that this is going to be the only thing asked on my next biology test, and I knew that I had to understand this or I'm doomed. This video helped so much!

[@raquelvargas487]

Thanks so much! I have an exam based on this lab and this really helped tie it all together.

[@surat8591]

Thank you for explaining everything in detail and actually articulating each word. I had a similar lab last week, but I didn't really understand the promoter and regulator part until now. Thank you! I wish you were my professor 😍😍

[@DocRonBio]

Glad to help!

[@__zelda__]

I have an exam tomorrow and this was so helpful! Thank you!!

[@DocRonBio]

Zelda Fogle Glad to help! Good luck on your exam!

[@FPtz]

Wish I had found this video before my lab. After many other videos everything was still unclear. After this video, i fully understand and can answer all of the post Lab questions!

[@PA30000]

Thanks, this was nice and simple, panic averted.

[@ecsl9796]

i had an emergency the week we discussed this in lab so i had no idea about this. I have my final in 4 days and this really helped me understand the whole lab :,) thank you!

[@pkre707]

Did this lab a few weeks ago. So much fun!

[@DanielWilliams-l4s]

Boehm Terrace

[@tennyvasghanian3015]

Such a clear explanation! Thank you!!

[@DocRonBio]

Tenny Vasghanian I appreciate the feedback. Glad I could help!

[@JeffreyChitwood-g1u]

Georgiana Flat

[@XTheSpartanX7]

wow this was great

[@TommyHugh-i5o]

Sylvester River

[@HumeRachel-j3w]

Glover Field

[@graycenamos1166]

super helpful!

[@peterl5337]

Amazing, you made it quite easy.

[@MichelleWolf-n5k]

Reinger Garden

[@cindyseymour5863]

Please consider correcting the captions! Extremely important for students with certain ADA accommodations. For example "pGlo" is automatically captioned as "pee glow" :-).

[@abigailb8102]

Thank you so much for this video!

[@alannaburns4725]

Amazing video!

[@DocRonBio]

Thank you! I appreciate it.

[@andyzzh8622]

Love this!Thank you soooo much!!

[@DocRonBio]

Happy to help!

[@DMorningStarWithAyandaSithole]

Thee best, thank you so much!!!❤

[@royarad1554]

Great video!

[@Avm2]

What is the independent variable?

[@thesavvybrand]

Thank You!

[@hillary7261]

Excellent! Thank you so much!

[@DocRonBio]

H M Glad it was helpful!

[@MohammedJalloh-v3y]

Great explanation!

[@evelynli2053]

this video was too good

[@sandisonkosi9835]

Doc Ron I thank you Sir

[@DocRonBio]

Glad to help! Good luck on the exam!

[@champ1177]

How would I use my knowledge of natural selection to explain why the LB+ plate didn't glow.

[@DocRonBio]

LB+ never received pGLO, thus they do not have the genes for that phenotype.

[@montruc8082]

Thank you ❤️

[@DocRonBio]

Mon Truc You are welcome!

[@iqrarafiq7065]

how do we know that LB is the positive control instead of being the negative control

[@DocRonBio]

Hello Iqra. In general, controls serve as a comparison or reference for your experiment. When the control serves to show the absence or loss of something we generally refer to that as a negative control. For example, when performing a chemical reaction of some kind, if you leave out a key reactant, showing that the reaction stops. In the case of pGlo, showing that without the plasmid the bacteria die because they do not have the gene for the antibiotic resistance trait. In the case of the positive control, we do this to show when some thing is working or present. For example, showing a previously confirmed functional reaction side by side with a new, experimental one. In the case of pGlo, we show that the bacteria are alive and able to be seen on the plate, even after heat shock. Otherwise, someone might argue that there never were bacteria to begin with in the experiment! Or if they are not present in the positive control you can show that there is something wrong with the bacteria to start. I hope this helps to clarify things!

[@iqrarafiq7065]

Doc Ron Bio thank you so much !

[@juelb4658]

do we get the same transformation efficiency for the last two plates? so for LB+amp and LB+amp+ara

[@DocRonBio]

Hello. Yes you should get roughly equal rates of transformation on these two plates (roughly 75 colonies). The plasmid being transformed is the same in both. The arabinose inducer is present on only one plate (LB+apm+ara), but his only impacts expression of GFP, not the transformation (which happens in a previous step). Transformation may, however, differ in each group and from plate to plate based on how well students follow the protocol (but not because of the molecular biology).

[@juelb4658]

@@DocRonBio That makes sense, thank you!

[@sophieni2931]

What does LB do?

[@DocRonBio]

The LB contains the nutrients the bacteria need to grow. So it is just their “food”.

[@sophieni2931]

@@DocRonBio thanks for responding!

[@champ1177]

What environmental selection took place in the petri dishes?

[@DocRonBio]

This would be antibiotic selection. If you do not have the resistance gene you are selected against.